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Journal of Southern Medical University ; (12): 1065-1070, 2019.
Article in Chinese | WPRIM | ID: wpr-773499

ABSTRACT

OBJECTIVE@#To isolate tumor stem-like cells from human epithelial ovarian cancer SKOV3 cells and explore their role in the formation of vascularization mimicry (VM).@*METHODS@#SKOV3 cells were passaged to the 7th generation by suspension culture in serum-free medium, and the percentages of CD133- and CD117-positive cells in the 1st, 3rd, 5th and 7th generations were analyzed using flow cytometry. The proliferative activity of the cells sorted from the 7th generation SKOV3 cells was assessed with colony formation assay. A three-dimensional cell culture model was established to compare the ability of VM formation between the sorted cells and the parental SKOV3 cells. The expression levels of matrix metalloproteinases-2 (MMP-2) and MMP-9 in the two groups were detected using real-time PCR and Western blotting.@*RESULTS@#Some SKOV3 cells formed typical cell spheres with suspension growth in serum-free medium and were passaged to the 7th generation. Flow cytometry revealed that the percentage of CD133-positive cells increased with cell passaging. The cloning efficiency of the sorted cells was significantly higher than that of the parental SKOV3 cells (50.33% 5.33%, < 0.001). The VM formation ability of the sorted cells was stronger than that of the parental SKOV3 cells in the three-dimensional cell culture system. RT-PCR and Western blotting showed that the expression levels of MMP-2 and MMP-9 were significantly higher in the 7th passage cells than in the parental cells ( < 0.05).@*CONCLUSIONS@#The sorted cells from SKOV3 cells cultured in serum-free medium exhibit biological properties of tumor stem cells with strong VM formation ability, suggesting their role in VM formation.


Subject(s)
Female , Humans , Carcinoma, Ovarian Epithelial , Pathology , Cell Line, Tumor , Cell Movement , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Neoplastic Stem Cells , Cell Biology , Neovascularization, Pathologic , Pathology , Ovarian Neoplasms , Pathology
2.
Chinese Journal of Clinical Oncology ; (24): 1124-1127, 2015.
Article in Chinese | WPRIM | ID: wpr-485116

ABSTRACT

Objective: To investigate the expression of vasculogenic mimicry (VM) in endometrial carcinoma tissues and its rela-tionship with the clinicopathologic features and prognosis of the disease. Methods: A total of 267 paraffin-embedded endometrial carci-noma specimens of patients with complete follow-up data were collected from the Shijiazhuang Bethune International Peace Hospital between January 2005 and June 2014. CD31-PAS dual staining was performed to identify the VM structure. Tissue samples were then divided into VM-positive and VM-negative groups. CD133 expression was detected by immunohistochemical streptavidin peroxidase method. Results: Among the 267 endometrial carcinoma patients, 65 cases (24.3%) were VM positive. VM formation was closely corre-lated with the International Federation of Gynecology and Obstetrics Staging (χ2=9.987, P=0.002), histodifferentiation grade (χ2=11.795, P=0.001), myometrial invasion depth (χ2=5.499, P=0.019), vascular cancer embolus (χ2=22.599, P<0.001), and lymph node me-tastasis (χ2=7.848, P=0.005). Kaplan-Meier survival curve analysis showed that survival time was significantly shorter in the VM-posi-tive group (median survival time was 51 months) than in the VM-negative group (median survival time was 100 months) (χ2=70.973, P<0.001). Moreover, CD133 expression was significantly higher in the VM-positive group [75.4% (49/65)] than in the VM-negative group [58.9% (119/202)] (χ2=5.720, P=0.017). Conclusion: VM is closely correlated with the pathogenesis, development, and malignancy of endometrial carcinoma. Furthermore, VM is one of the important indexes influencing the prognosis of this disease. Therefore, CD133-positive cells may contribute to VM formation.

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